Schröpfer Susan, Lempe Janne, Emeriewen Ofere Francis, Flachowsky Henryk
Julius Kühn Institute (JKI) - Federal Research Centre for Cultivated Plants, Institute for Breeding Research on Fruit Crops, Dresden, Germany.
Front Plant Sci. 2022 Jun 30;13:928292. doi: 10.3389/fpls.2022.928292. eCollection 2022.
Genetic transformation has become an important tool in plant genome research over the last three decades. This applies not only to model plants such as but also increasingly to cultivated plants, where the establishment of transformation methods could still pose many problems. One of such plants is the apple ( spp.), the most important fruit of the temperate climate zone. Although the genetic transformation of apple using has been possible since 1989, only a few research groups worldwide have successfully applied this technology, and efficiency remains poor. Nevertheless, there have been some developments, especially in recent years, which allowed for the expansion of the toolbox of breeders and breeding researchers. This review article attempts to summarize recent developments in the -mediated transformation strategies of apple. In addition to the use of different tissues and media for transformation, agroinfiltration, as well as pre-transformation with a Baby boom transcription factor are notable successes that have improved transformation efficiency in apple. Further, we highlight targeted gene silencing applications. Besides the classical strategies of RNAi-based silencing by stable transformation with hairpin gene constructs, optimized protocols for virus-induced gene silencing (VIGS) and artificial micro RNAs (amiRNAs) have emerged as powerful technologies for silencing genes of interest. Success has also been achieved in establishing methods for targeted genome editing (GE). For example, it was recently possible for the first time to generate a homohistont GE line into which a biallelic mutation was specifically inserted in a target gene. In addition to these methods, which are primarily aimed at increasing transformation efficiency, improving the precision of genetic modification and reducing the time required, methods are also discussed in which genetically modified plants are used for breeding purposes. In particular, the current state of the rapid crop cycle breeding system and its applications will be presented.
在过去三十年里，遗传转化已成为植物基因组研究中的一项重要工具。这不仅适用于诸如 等模式植物，也越来越多地适用于栽培植物，而栽培植物转化方法的建立仍可能存在许多问题。苹果（ spp.）就是这样一种植物，它是温带气候区最重要的水果。尽管自1989年以来就有可能利用 对苹果进行遗传转化，但全球只有少数研究小组成功应用了这项技术，而且效率仍然很低。然而，尤其是近年来已经有了一些进展，这使得育种者和育种研究人员的工具库得以扩展。这篇综述文章试图总结苹果 介导转化策略的最新进展。除了使用不同的组织和培养基进行转化外，农杆菌浸润以及用婴儿潮转录因子进行转化前处理都是显著的成功案例，它们提高了苹果的转化效率。此外，我们还强调了靶向基因沉默的应用。除了通过用发夹基因构建体进行稳定转化实现基于RNAi的经典沉默策略外，病毒诱导基因沉默（VIGS）和人工微小RNA（amiRNA）的优化方案已成为沉默目标基因的强大技术。在建立靶向基因组编辑（GE）方法方面也取得了成功。例如，最近首次有可能产生一个同源组GE系，其中在目标基因中特异性插入了双等位基因突变。除了这些主要旨在提高转化效率、提高基因修饰精度和减少所需时间的方法外，还讨论了将转基因植物用于育种目的的方法。特别是，将介绍快速作物周期育种系统的现状及其应用。
